[Effects of mild moxibustion on visceral hypersensitivity via SCF/c-kit signaling pathway in diarrhea-predominant irritable bowel syndrome rats with syndrome of liver-qi stagnation and spleen deficiency]. / åŸºäºŽå¹²ç»†èƒžç”Ÿé•¿å› å­/é ªæ°¨é ¸æ¿€é ¶å—ä½“ä¿¡å·é€šè·¯æŽ¢è®¨æ¸©å’Œç¸å¯¹è‚éƒè„¾è™šè¯è ¹æ³»åž‹è‚ æ˜“æ¿€ç»¼åˆå¾å¤§é¼ å† è„é«˜æ•æ„Ÿçš„å½±å“.

OBJECTIVES: To observe the effect of mild moxibustion on stem cell factor (SCF)/tyrosine kinase receptor (c-kit) signaling pathway and visceral hypersensitivity of diarrhea predominant irritable bowel syndrome (IBS-D) model rats with liver-qi stagnation and spleen deficiency syndrome, so as to explore its mechanisms underlying improvement of IBS-D. METHODS: A total of 24 male Wistar rats were randomly divided into normal, IBS-D model, medication and mild moxibustion groups, with 6 rats in each group. The IBS-D model was established by glacial acetic acid (4%) enema plus restraint stress stimulation once daily for 14 days. Rats of the medication group were treated by gavage of pivamium bromide (15 mg/kg) once a day for 14 days. Mild moxibustion was applied to bilateral "Tianshu"(ST25), "Shangjuxu"(ST37) and "Taichong"(LR3) for 20 min, once daily for 14 consecutive days. After the intervention, the rats' general state of each group were observed. The rate of loose stools (LSR), and the minimum volume threshold for abdominal withdrawal reflex(AWR) were observed, and the open field test was used to assess the state of rats' motor activities (including rearing times, grooming times and total number of square-crossings in 5 min). Morphological changes of the colon tissue were observed by hematoxylin-eosin (H.E.) staining, The count of mast cells (MC) in the colon tissues was determined by toluidine blue staining. Contents of serum 5-hydroxytryptamine (5-HT) and substance P (SP) were determined by enzyme-linked immunosorbent assay (ELISA). The relative expression levels of SCF and c-kit mRNAs and proteins in the colon tissues were detected by real-time quantitative PCR and Western blot, respectively. RESULTS: Compared with the normal group, the body weight, minimum volume threshold of AWR, total numbers of square-crossing, rearing times and grooming times were significantly decreased (P<0.05), and the LSR, number of MC, contents of 5-HT and SP, and the expression levels of SCF and c-kit mRNAs and proteins were considerably increased in the model group (P<0.01). In comparison with the model group after interventions, the body weight, minimum volume threshold of AWR, total numbers of square-crossing, rearing times and grooming times were apparently increased in both medication and moxibustion groups (P<0.05, P<0.01), and the LSR, number of MC, 5-HT and SP contents in both medication and moxibustion groups, and the expression levels of SCF and c-kit mRNA and protein in the moxibustion group (not in the medication group) were obviously decreased (P<0.05, P<0.01). H.E. staining showed that in the model group, a small amount of inflammatory cells in the mucosal layer of colon tissue could be seen. in the medication group, a small number of lymphocytes in colon tissue were observed, while in the mild moxibustion group, a small amount of neutrophils in colon tissue were observed. CONCLUSIONS: Mild moxibustion can reduce visceral hypersensitivity and improve abdominal pain, diarrhea and locomotion state in IBS-D rats with liver-qi stagnation and spleen deficiency syndrome, which may be associated with its functions in reducing the number of MC and the levels of 5-HT and SP and down-regulating the activities of SCF/c-kit signaling pathway.

Effect of moxibustion on the SIRT1/FOXO3a signaling pathway in ApoE-/- mice with atherosclerosis. / 艾灸对ApoE-/-åŠ¨è„‰ç²¥æ ·ç¡¬åŒ–å°é¼ SIRT1/FOXO3a信号通路的影响.

OBJECTIVES: To observe the effects of moxibustion on blood lipid metabolism, pathological morphology of thoracic aorta, and the expression of silent information regulator 1 (SIRT1) and forkhead box transcription factor O3a (FOXO3a) in ApoE-/- atherosclerosis (AS) mice, so as to explore the potential mechanism of moxibustion in preventing and treating AS. METHODS: Ten C57BL/6J mice were fed a normal diet as the control group, and 30 ApoE-/- mice were fed a high-fat diet to establish the AS model, which were randomly divided into the model group, simvastatin group, and moxibustion group, with 10 mice in each group. From the first day of modeling, mice in the moxibustion group received mild moxibustion treatment at "Shenque"(CV8), "Yinlingquan"(SP9), bilateral "Neiguan"(PC6) and "Xuehai"(SP10) for 30 min per time；the mice in the simvastatin group were given simvastatin orally (2.5 mg·kg-1·d-1), with both treatments given once daily, 5 times a week, with a total intervention period of 12 weeks. The body weight and general condition of the mice were observed and recorded during the intervention period. After the intervention, the contents of serum total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) were measured using an automated biochemistry analyzer. Hematoxylin eosin (HE) staining was used to observe the pathological morphology of the thoracic aorta. ELISA was used to measure the contents of serum oxidized low-density lipoprotein (ox-LDL) and superoxide dismutase (SOD) activity. Western blot and real-time fluorescent quantitative PCR analysis were used to detect the expression levels of SIRT1 and FOXO3a protein and mRNA in the thoracic aorta. RESULTS: Compared with the control group, body weight at the 8th and 12th week, serum TC, TG, LDL-C, and ox-LDL contents of the model group mice were significantly increased(P<0.05, P<0.01), while the HDL-C contents, SOD activity, and the expression levels of SIRT1 protein and mRNA in the thoracic aorta were significantly decreased(P<0.05, P<0.01). HE staining showed thickening of the aortic intima, endothelial cell degeneration, swelling, and shedding. Compared with the model group, body weight at the 8th and 12th week, serum TC, TG, LDL-C, and ox-LDL contents of mice in the simvastatin group and moxibustion group were significantly decreased(P<0.01), while the serum SOD activity, expression levels of SIRT1 protein and mRNA in the thoracic aorta were significantly increased(P<0.01). The HDL-C contents were significantly increased in the simvastatin group(P<0.05). The thoracic aortic structure was more intact in both groups, with a more regular lumen and orderly arrangement of the elastic membrane in the media, and a slight amount of endothelial cell degeneration and swelling in the intima. There was no significant difference in the evaluated indexes between the moxibustion group and the simvastatin group and the pathological changes in the thoracic aorta were similar between the two groups. CONCLUSIONS: Moxibustion can reduce the body weight of AS model mice, regulate lipid levels, repair vascular intima, and alleviate endothelial damage. Its mechanism of action may be related to the regulation of the SIRT1/FOXO3a signaling pathway to improve oxidative damage.

Effect of acupoint catgut embedding on p38 MAPK pathway in the lung tissue of asthmatic rats. / 穴位埋线通过调节p38ä¸è£‚åŽŸæ´»åŒ–è›‹ç™½æ¿€é ¶é€šè·¯å¯¹å“®å–˜å¤§é¼ æ°”é“ä¸Šçš®çš„å½±å“.

OBJECTIVES: To observe the effect of catgut embedding at "Feishu"(BL13), "Dingchuan" (EX-B1) and "Danzhong" (CV17) on expression of phosphorylated p38 mitogen activated protein kinase (p-p38 MAPK), interleukin-4 (IL-4), interferon-γ (IFN-γ) and changes of airway epithelial cells (AEC) in the lung tissue of bronchial asthma (BA) rats, so as to explore its mechanisms underlying improvement of BA. METHODS: Forty male Wistar rats were randomly and equally divided into blank control, model, dexamethasone (DEX) and catgut embedding groups. The BA model was established by intraperitoneal injection of suspension of ovalbumin and aluminum hydroxide. Rats of the DEX group received intraperitoneal injection of DEX (1.5 mg/kg), once daily for 2 weeks, and those of the catgut embedding group received catgut embedding at BL13, EX-B1 and CV17 only one time. The rats' sneezing times per miniute in each group were recorded. H.E. staining was used to observe the histopathological changes of the lung tissue under light microscope. A transmission electron microscope (TEM) was used to observe the ultrastructural changes of AEC in the lung tissue, including the thickness of bronchial wall and bronchial smooth muscle by using an image analysis software. The protein expressions of p-p38 MAPK, IL-4 and INF-γ in the lung tissue were determined using Western blot. RESULTS: Morphological observation revealed that in the model group, light microscope showed deformed and swollen bronchial tube wall with increased folds and thickened bronchial smooth muscle；and TEM showed a large number of autophagy vesicles containing swollen and deformed organelles in the AEC, and apparent reduction of intracellular mitochondria, these situations were obviously milder in both DEX and catgut embedding groups. Compared with the blank control group, the sneezing times, thickness of bronchial wall and bronchial smooth muscle in the model group were significantly increased (P<0.01), and the expressions of p-p38 MAPK and IL-4 in lung tissue were significantly increased (P<0.01), while the expression of IFN-γ was significantly decreased (P<0.01) in the model group. In comparison with the model group, the sneezing times, thickness of bronchial wall and bronchial smooth muscle, protein expressions of p-p38 MAPK and IL-4 were significantly decreased (P<0.01), while the expression of IFN-γ was obviously increased (P<0.01) in both the DEX and catgut embedding groups. CONCLUSIONS: Acupoint catgut embedding can reduce the expression of IL-4 and increase the expression of IFN-γ by inhibiting p38 MAPK signal pathway of lung tissues in BA rats, which may contribute to its effect in alleviating the degree of airway epithelial cells damage.

Effect of acupuncture on the expression of neuropeptides and related inflammatory factors in rats with diarrhea-predominant irritable bowel syndrome. / é’ˆåˆºå¯¹è ¹æ³»åž‹è‚ æ˜“æ¿€ç»¼åˆå¾å¤§é¼ è„‘è‚ è‚½åŠç›¸å ³ç‚Žæ€§å› å­è¡¨è¾¾çš„å½±å“.

OBJECTIVES: To observe the effect of acupuncture on the expressions of neuropeptides and related inflammatory factors in rats with diarrhea-predominant irritable bowel syndrome(IBS-D), so as to explore the mechanism of acupuncture in the treatment of IBS-D. METHODS: Male Wistar rats were randomly divided into blank group, model group, medication group, and acupuncture group, with 6 rats in each group. Except for the blank group, the other groups were subjected to 14-day "acetic acid enema + restraint stress" to establish the IBS-D rat model. After successful modeling, the medication group received gavage of pinaverium bromide(15 mg/kg) once a day, and the acupuncture group received acupuncture at "Baihui"(GV20) and bilateral "Tianshu"(ST25), "Shangjuxu"(ST37), "Zusanli"(ST36), and "Taichong"(LR3) for 20 min every day, both groups were treated continuously for 14 days. The general state of the rats in each group was observed, and the body weight of the rats was measured. The open-field experiment was conducted to measure the horizontal and vertical movements, and the number of fecal pellets of rats. The histopathological morphology of hypothalamus and colon of rats was observed by HE staining. Toluidine blue staining was used to observe and count the mast cells(MCs) in the colon tissue of rats. ELISA was used to detect the serum contents of tumor necrosis factor-α(TNF-α) and interleukin(IL)-10. Real-time fluorescence quantitative PCR was performed to detect the mRNA expressions of calcitonin gene-related peptide(CGRP) in the hypothalamus and colon tissue. Western blot was used to detect the expressions of corticotropin-releasing factor(CRF) in the hypothalamus and colon tissue. RESULTS: HE staining showed that there was inflammatory cell infiltration in the lamina propria of colon in the model group, and it was reduced in the other groups. Compared with the blank group, the model group showed significantly decreased body weight, decreased walking distance and upright times in open field experiment, decreased serum IL-10 contents(P<0.05, P<0.01), increased fecal pellet number (P<0.01), increased MC number in the colon tissue, serum TNF-α contents, and CGRP mRNA expressions and CRF expressions in the hypothalamus and colon tissue(P<0.01). Compared with the model group, both medication and acupuncture groups showed significantly increased body weight, walking distance and upright times in the open-field experiment, and serum IL-10 contents(P<0.01, P<0.05), significantly decreased fecal pellet number (P<0.05), significantly decreased MC number in the colon tissue, serum TNF-α contents, and CGRP mRNA expressions in the hypothalamus and colon tissue(P<0.01)；at the same time, the acupuncture group showed significantly decreased CRF expressions in the hypothalamus and colon tissue(P<0.01, P<0.05). There was no significant difference in the above indicators between the medication group and the acupuncture group. CONCLUSIONS: Acupuncture can improve the general and emotional state, inflammatory response, and neuropeptide expression in rats with IBS-D, and alleviate the symptoms of IBS-D, which may be related to the regulation of neuropeptides and inflammatory factors levels.